Planta Med 2008; 74 - PG54
DOI: 10.1055/s-0028-1084806

Establishment of Rindera graeca transgenic root culture as a source of shikonin derivatives

K Sykłowska-Baranek 1, A Pietrosiuk 1, Ł Kuźma 2, I Chinou 3, M Kongel 1, M Jeziorek 1
  • 1Department of Biology and Pharmaceutical Botany, Warsaw Medical University, Banacha 1, 02–097 Warsaw, Poland
  • 2Department of Biology and Pharmaceutical Botany, Medical University of Łódź, Muszyńskiego 1, 90–151 Łódź, Poland
  • 3Department of Pharmacognosy, School of Pharmacy, University of Athens, University Campus of Zografou, 157 71 Zografou Athens, Greece

Rindera graeca Boiss. & Heldr. (Boraginaceae) is an endemic plant of South-East Europe and Mediterranean Basin, growing in rocky places of Greece. The preliminary chemical investigation showed the presence of shikonin derivatives in the natural roots of this species cultured in vitro. The inoculation of four-week-old plantlets of R.graeca with Agrobacterium rhizogenes ATCC 15834 strain resulted in 60 transgenic root lines. For the further investigation 4 of them were chosen: KTRg3, KTRg7, KTRg17 and KTRg18. The hairy roots were cultivated in Erlenmeyer flasks containing liquid hormone-free media LS [1], DCR [2] and three different modification of DCR medium. The line KTRg7 was also cultivated in a sprinkle bioreactor where roots grew in LS and next in M9 [3] medium. The highest biomass increase was observed during cultivating roots of KTRg18 line in DCR –62-fold. The roots of KTRg7 line maintained in LS medium in bioreactor demonstrated mild growth and turned brown after 2 weeks from the beginning of the culture. After medium changing to M9 (on 35 day) the hairy roots grew intensively.

The transgenic roots collected from different media were investigated using HPLC UV-DAD method. In all examined hairy root lines there was no shikonin. On the basis of UV-spectra four shikonin derivatives were detected in roots cultivated in modified DCR medium.

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